In:
Cell Proliferation, Wiley, Vol. 48, No. 6 ( 2015-12), p. 729-737
Abstract:
This study aimed to investigate the role of the TGF β signalling pathway in angiogenesis in a three‐dimensional (3D) collagen gel model, with co‐culture between adipose‐derived stromal cells ( ASC s) and endothelial cells ( EC s). Materials and methods A 3D collagen gel, implanted with green fluorescent protein‐labelled mouse ASC s and red fluorescent protein‐labelled mouse EC s, was established in vitro . Phenomena of angiogenesis with or without type I TGF β receptor inhibitor ( LY 2157299) treatment, were observed 7 days post‐implantation, using confocal laser scanning microscopy. To detect expression of angiogenesis‐related genes, semi‐quantitative PCR and quantitative real‐time PCR were conducted. Zymography was performed to explore secretion of matrix metalloproteinase 2 ( MMP ‐2) and matrix metalloproteinase 9 ( MMP ‐9) after treatment with LY 2157299 of 5, 10, 20 to 50 μ m concentrations, for 24 h. Results Angiogenesis was found to be attenuated in co‐culture gels after ASC and EC treatment with LY 2157299. Genes VEGF ‐A , VEGF ‐B , VE ‐ca , FGF ‐1 , FGF ‐2 , PDGF , HGF , BMP ‐4 were significantly reduced in the presence of LY 2157299 in both mono‐cultured and co‐cultured EC s. Furthermore, reduction in co‐cultured EC s was prominent relative to mono‐cultured EC s, while the same results did not occur to ASC s. We further confirmed that gelatinases secreted by EC s were reduced in a dose‐dependent manner, after treatment with LY 2157299. Conclusions These results indicate that in ASC / EC co‐culture, the TGF β signalling pathway regulated angiogenesis via EC activity. Co‐cultured EC s were regulated more significantly than mono‐cultured EC s suggesting that inhibition of TGF β RI may regulate paracrine secretion of ASC s to further modulate EC angiogenesis.
Type of Medium:
Online Resource
ISSN:
0960-7722
,
1365-2184
DOI:
10.1111/cpr.2015.48.issue-6
Language:
English
Publisher:
Wiley
Publication Date:
2015
detail.hit.zdb_id:
2019986-7
SSG:
12
Bookmarklink